R25 Advanced Techniques in Single Cell Transcriptomics (J. Eberwine/J. Kim) Project Summary The Penn Genome Frontiers Institute (PGFI) will offer an Advanced Techniques in Single Cell Transcriptomics five-day course in 2014, 2015 and 2016 that would be open to researchers (PIs, Postdocs, and Graduate Students) nationally. Course participants will be trained to successfully perform the entire process of quantifying RNA from individual cells from multi- cellular organisms. The base set of techniques that will be taught include single cell isolation, single cell RNA isolation, RNA amplification by aRNA and PCR, NextGen seq library construction, sequencing and data analysis. Lectures and discussions on the broader context of single cell transcriptomics within single cell analysis and on the most recent technological developments will enhance the hands-on bench training. The final session of the course will cover RNAseq data analysis and interpretation. Course materials, including basic curriculum, protocols and datasets, will be available on PGFI-hosted, publically accessible web pages. This Single Cell Transcriptomics course will provide a research training opportunity for which we expect there to be broad interest and applicability among researchers working in gene expression and functional genomics in a diverse range of fundamental and biomedical disciplines. Researchers constrained by access to very small cell populations or directly interested in cell heterogeneity (e.g., characterization of transcriptomes, development and normal states, cancer and other disease-state genomics) and its consequences would gain valuable tools from this workshop. Dissemination of tools for single cell analysis is critical for enabling this promising area to expand and deliver on its potential.